To obtain ultrafiltration (UF) membranes with excellent separation performances and antifouling properties, regenerated cellulose (RC) membranes with intrinsic hydrophilicity and dense structures immensely attract the attention of researchers. Herein, a novel modified RC membrane, which has high water permeability and antifouling capability, is prepared using cellulose acetate (CA) via non-solvent induced phase separation (NIPS) and green hydrolysis technology. The as-prepared membranes were investigated comprehensively by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) analyses and protein adsorption, etc. As a result, the RC membranes showed higher hydrophilicity (WCA=25.95°), thus they possessed a lower protein adsorption value of 1.9 ± 0.1 μg/cm². The filtration performances were evaluated by bovine serum albumin (BSA) and endotoxin separation with dead-end filtration system. The results exhibited greater selectivity of RC membranes that BSA rejection over 98% and endotoxin remove below 0.25 EU/mL, and excellent anti-fouling properties that the flux recovery rate (FRR) over 95% and even as high as 100%. Simultaneously, the common polyethersulfone (PES) composite membranes were prepared via NIPS technology, and comprehensively compared with RC membranes to provide a theoretical basis for the practical application of RC membranes in pharmaceutical and biotechnological industries.